A method of a bicolor fast-Fourier pulse-amplitude modulation chlorophyll fluorometry


A simple method of a bicolor (multicolor), fast-Fourier, PAM chlorophyll fluorometry has been developed to obtain fluorescence induction curves. Quantum yields of PSII photochemistry were determined with blue and red simultaneously applied pulsed measuring lights for three subsequent 20-min periods of dark-, light-adaptation under actinic light and dark recovery. Measuring lights were cross-combined with blue and red actinic lights and saturation pulses. Coefficients of chromatic divergence were calculated as a ratio of the quantum yields obtained by red measuring light to that obtained by blue measuring light. Adaptation of Ficus benjamina and Hordeum vulgare leaves under blue (but not red) actinic light resulted in the sufficient increase of chromatic divergence. In addition, fraction of active, non(photo)inhibited, PSII centers was shown to be dependent on the color of measuring light. Thus, color of the light sources should be considered when reporting results of parameters evaluated from fluorescence induction curves.

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AB and AR:

blue and red actinic lights


actinic light


operating or effective Chl fluorescence level in the light-adapted state

F0 :

minimal Chl fluorescence level in dark-adapted state


fast Fourier transform

Fm, Fm' :

maximal Chl fluorescence level in dark-adapted and light-adapted state

Fm1'' and Fm2'' :

maximal Chl fluorescence levels recovered at the beginning and at the end of the second dark period

Fv :

variable Chl fluorescence


light-emitting diode

MB and MR:

blue and red measuring flashes


measuring light


saturating pulse


pulse amplitude modulation (fluorometry)

SB and SR:

blue and red saturating pulses


chromatic divergence of quantum yields of PSII photochemistry


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Correspondence to V. Lysenko.

Additional information

Acknowledgements: V.L. and T.V. were supported by Ministry of Education and Science of Russian Federatian by grant BCh0110- 11/2017-25 and D.L. was supported by grant LO1204 from the National Program of Sustainability I, Ministry of Education, Youth and Sports, Czech Republic. The laboratory experiments and measurements were supported by grant BCh0110-11/2017-25 and performed with the equipment of Multiaccess center “Biotechnology, Biomedicine and Environmental Monitoring”, Laboratory of Plant Physiology and Ecology of Academy of Biology and Biotechnology of Southern Federal University.

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Lysenko, V., Lazár, D. & Varduny, T. A method of a bicolor fast-Fourier pulse-amplitude modulation chlorophyll fluorometry. Photosynthetica 56, 1447–1452 (2018). https://doi.org/10.1007/s11099-018-0848-y

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Additional key words

  • chromatic adaptation
  • greenhouse lighting
  • dual-wavelength PAM
  • multi-color PAM